1H-pyrido[2,3-b][1,5]benzodiazipine vasopressin agonists

ABSTRACT

The present invention provides compounds of the general formula:                    
     wherein Y is a moiety selected independently, from NH or —(CH 2 ) n — wherein n is 1; m is an integer from 1 to 2; and the moiety                    
     represents: (1) a 5-membered aromatic (unsaturated) heterocyclic ring having one nitrogen (wherein A is nitrogen, and B and C are CH), (2) a 6-membered aromatic (unsaturated) heterocyclic ring having one nitrogen (wherein A is carbon, B is nitrogen, and C is —CH—CH—), (3) a 6-membered aromatic (unsaturated) ring (wherein A is carbon, B is CH, and C is —CH—CH—); as well as methods and pharmaceutical compositions utilizing these compounds for the treatment of disorder which may be remedied or alleviated by vasopressin agonist activity, including diabetes insipidus, nocturnal enuresis, nocturia, urinary incontinence, bleeding and coagulation disorders, or temporary delay of urination.

This application is a division of Ser. No. 09/495,505 filed Feb. 1, 2000and also claims the benefit of U.S. Provisional Application No.60/155,235, which was converted from U.S. patent application Ser. No.09/244,179, filed Feb. 4, 1999, pursuant to a petition filed under 37C.F.R. 1.53(c)(2)(i).

This invention concerns tricyclic arylthiophene compounds which act asvasopressin V₂ agonists, as well as methods of treatment andpharmaceutical compositions utilizing these compounds.

BACKGROUND OF THE INVENTION

Vasopressin (antidiuretic hormone, ADH) a nonapeptide hormone andneurotransmitter, is synthesized in the supraoptic nuclei of thehypothalamus of the brain and transported through thesupraoptico-hypophyseal tract to the posterior pituitary where it isstored. Upon sensing an increase in plasma osmolality by brainosmoreceptors or a decrease in blood volume or blood pressure (detectedby the baroreceptors and volume receptors), vasopressin is released intothe blood circulation and activates vasopressin V_(1a) receptors onblood vessels causing vasoconstriction to raise blood pressure; andvasopressin V₂ receptors of the nephron of the kidney causingreabsorption mainly of water and to a lesser degree electrolytes, toexpand the blood volume (Cervoni and Chan, Diuretic Agents, inKirk-Othmer, Encyclopedia of Chemical Technology, 4th ed., Wiley, Volume8, 398-432, (1993)). The existence of vasopressin in the pituitary wasknown as early as 1895 (Oliver and Schaefer, J. Physiol. (London), 18,277-279, (1895)). The determination of the structure and the totalsynthesis of vasopressin were accomplished by du Vigneaud and coworkersin 1954 (du Vigneaud, Gish and Katsoyannis, J. Am. Chem. Soc., 76,4751-4752, (1954)).

The actions of vasopressin V_(1a) receptors are mediated through thehosphatidylinositol pathway. Activation of vasopressin V_(1a) receptorscauses contraction of the smooth muscle of the blood vessels to raiseblood pressure. The actions of the vasopressin V₂ receptors are mediatedthrough activation of the denylate cyclase system and elevation ofintracellular levels of cAMP. The activation of vasopressin V₂ receptorsby vasopressin or vasopressin-like (peptide or non-peptidic) compoundsincreases water permeability of the collecting ducts of the nephron andpermits the reabsorption of a large quantity of free water. The endresult is the formation and excretion of a concentrated urine, with adecrease in urine volume and an increase in urinary osmolality.

Vasopressin plays a vital role in the conservation of water byconcentrating the urine at the site of the collecting ducts of thekidney. The collecting ducts of the kidney are relatively impermeable towater without the presence of vasopressin at the receptors andtherefore, the hypotonic fluid formed after filtering through theglomeruli, passing the proximal convoluted tubule, the loops of Henle,and the distal convoluted tubules, will be excreted as dilute urine.However, during dehydration, volume depletion or blood loss, vasopressinis released from the brain and activates the vasopressin V₂ receptors inthe collecting ducts of the kidney rendering the ducts very permeable towater; hence water is reabsorbed and a concentrated urine is excreted.In patients and animals with central or neurogenic diabetes insipidus,the synthesis of vasopressin in the brain is defective and therefore,they produce very little or no vasopressin, but their vasopressinreceptors in the kidneys are normal. Because they cannot concentrate theurine, they may produce as much as 10 times the urine volumes of theirhealthy counterparts and they are very sensitive to the action ofvasopressin and vasopressin V₂ agonists. Vasopressin and desmopressin,which is a peptide analog of the natural vasopressin, are being used inpatients with central diabetes insipidus. Vasopressin V₂ agonists arealso useful for the treatment of nocturnal enuresis, nocturia, urinaryincontinence and temporary delay of urination whenever desirable.

Vasopressin, through activation of its V_(1a) receptors, exertsvasoconstricting effects so as to raise blood pressure. A vasopressinV_(1a) receptor antagonist will counteract this effect. Vasopressin andvasopressin-like agonists release factor VIII and von Willebrand factorso they are useful for the treatment of bleeding disorders, such ashemophilia. Vasopressin and vasopressin-like agonists also releasetissue-type plasminogen activator (t-PA) into the blood circulation sothey are useful in dissolving blood clots such as in patients withmyocardial infarction and other thromboembolic disorders (Jackson,“Vasopressin and other agents affecting the renal conservation ofwater”, in Goodman and Gilman, The Pharmacological Basis ofTherapeutics, 9th ed., Hadman, Limbird, Molinoff, Ruddon and GilmanEds., McGraw-Hill, New York, pp. 715-731 (1996); Lethagen, Ann. Hematol.69, 173-180 (1994); Cash et al., Brit. J. Haematol., 27, 363-364 (1974);David, Regulatory Peptides, 45, 311-317 (1993); Burggraaf et al., Cli.Sci., 86, 497-503 (1994)).

The following prior art references describe peptidic vasopressinantagonists: Manning et al., J. Med. Chem., 35, 382 (1992); Manning etal., J. Med. Chem., 35, 3895 (1992); Gavras and Lammek, U.S. Pat. No.5,070,187 (1991); Manning and Sawyer, U.S. Pat. No. 5,055,448 (1991);Ali, U.S. Pat. No. 4,766,108 (1988); Ruffolo et al., Drug News andPerspectives 4(4), 217 (May 1991); Albright and Chan, Curr. Pharm. Des.3(6), 615 (1997). Williams et al., have reported on potent hexapeptideoxytocin antagonists [J. Med. Chem., 35, 3905 (1992)] which also exhibitweak vasopressin antagonistic activity in binding to V₁ and V₂receptors. Peptidic vasopressin antagonists suffer from a lack of oralactivity and many of these peptides are non-selective antagonists sincethey also exhibit partial agonist activity.

Non-peptidic vasopressin antagonists have recently been disclosed.Albright et al. describe tricyclic azepines as vasopressin antagonistsor vasopressin and oxytocin antagonists in U.S. Pat. No. 5,516,774(1996), U.S. Pat. No. 5,532,235 (1996), U.S. Pat. No. 5,536,718, U.S.Pat. No. 5, 610,156 (1997), U.S. Pat. No. 5,612,334 (1997), U.S. Pat.No. 5,624,923 (1997), U.S. Pat. No. 5,654,297 (1997), U.S. Pat. No.5,686,445 (1997), U.S. Pat. No. 5,693,635 (1997), U.S. Pat. No.5,696,112, U.S. Pat. No. 5,700,796 (1997), U.S. Pat. No. 5,719, 278(1998), U.S. Pat. No. 5,733, 905 (1998), U.S. Pat. No. 5,736,538 (1998),U.S. Pat. No. 5,736,540 (1998), U.S. Pat. No. 5,739,128 (1998), U.S.Pat. No. 5,747,487 (1998), U.S. Pat. No. 5,753,648 (1998), U.S. Pat. No.5,760,031 (1998), U.S. Pat. No. 5,780,471 (1998);tetrahydrobenzodiazepine derivatives as vasopressin antagonists aredisclosed in J.P. 0801460-A (1996); Ogawa et al., disclosebenzoheterocyclic derivatives as vasopressin and oxytocin antagonists,and as vasopressin agonists in WO 9534540-A; and Venkatesan et al.,disclose tricyclic benzazepine derivatives as vasopressin and oxytocinantagonists in U.S. Pat. No. 5,521,173 (1996).

As mentioned above, desmopressin (1-desamino-8-D-arginine vasopressin)(Huguenin and Boissonnas, Helv. Chim. Acta, 49, 695 (1966)) is avasopressin agonist. The compound is a synthetic peptide with variablebioavailability. An intranasal route is poorly tolerated and an oralformulation for nocturnal enuresis requires a 10-20 fold greater dosethan the intranasal administration.

Albright et al. disclose (cf. examples 1 and 6), a subset of tricyclicpyrrolo benzodiazepines which are part of the present application, as V₁and/or V₂ vasopressin receptor antagonists and oxytocin receptorantagonists in U.S. Pat. No. 5,521,173 (1996), initer alica.

Compounds of general structure 7a in Scheme I U.S. Pat. No. 5,521,173are taught by Albright et al. to possess antagonist activity at V₁and/or V₂ receptors and exhibit in vivo vasopressin antagonist activity,as well as antagonist activity at oxytocin receptors.

7a, Scheme I (Albright et al.) U.S. Pat. No. 5,521,173

wherein m is 1; Y is a moiety selected from (CH₂)_(n) wherein n is 1; D,E, and F are selected from carbon; A¹ is CH; R⁶ is the moiety:

K¹ is CH; X is S; R⁵ is hydrogen; R⁷ is hydrogen; and the moiety:

represents a fused phenyl or optionally substituted phenyl.

Also, Albright et al. broadly disclose a subset of tricyclic pyrrolo andpyrido benzodiazepines, part of the present application, as V₁ and/or V₂vasopressin receptor antagonists and oxytocin receptor antagonists in WO96/22282 A1 (1996), inter alia.

Compounds of general structure 61b in Scheme 12 of the aboveapplication, are claimed by Albright et al. to possess antagonistactivity at V₁ and/or V₂ receptors and exhibit in vivo vasopressinantagonist activity, as well as antagonist activity at oxytocinreceptors.

61b, Scheme 12 (Albright et al.) WO 96/22282 A1

wherein m is 1; Y is a NH or a moiety selected from (CH₂)_(n) wherein nis 1; R⁵ and R⁷ are selected from hydrogen; X is direct bond; R¹⁰represents the moiety

p is 0; R¹ and R² are selected from hydrogen, (C₁-C₃) lower alkyl,(C₁-C₃) lower alkoxy and halogen; and the moiety

represents an optionally substituted phenyl, a 5-membered aromatic(unsaturated) heterocyclic ring having one nitrogen atom, or a6-membered aromatic (unsaturated) heterocyclic ring having one nitrogenatom.

However, certain tricyclic pyrrolo- and pyridobenzodiazepines of generalstructure 7b and 61b have been found unexpectedly to be vasopressin V₂receptor agonists in vivo, and thus possess different biological profileand clinical utility from those originally disclosed. Thus, rather thanhaving an aquaretic effect they do unexpectedly cause reabsorption ofwater, i.e. they reduce urine volume and increase urine osmolality.

The compounds of this invention are non-peptidic and have a good oralbioavailability. They are vasopressin V₂ receptor agonists, and as suchthey promote reabsorption of water. They have no vasopressin V_(1a)receptor agonist effects so they do not raise blood pressure. Incontrast, the prior art compounds are described as vasopressinantagonists at both the V_(1a) and V₂ receptors.

SUMMARY OF THE INVENTION

This invention relates to novel and known compounds selected from thoseof formula (I):

wherein:

Y is a moiety selected independently, from NH or —(CH₂)_(n)— wherein nis 1; m is an integer from 1 to 2;

R¹, R², R⁵ and R⁶ are independently, selected from hydrogen, lower alkyl(C₁-C₆), lower alkoxy (C₁-C₆), halogen, and CF₃;

R₃ and R₄ are independently, selected from the group comprisinghydrogen, lower alkyl (C₁-C₆), halogen, amino, (C₁-C₆) lower alkoxy, or(C₁-C₆) lower alkylamino; and the moiety

represents:

(1) a 5-membered aromatic (unsaturated) heterocyclic ring having onenitrogen (wherein A is nitrogen, and B and C are CH);

(2) a 6-membered aromatic (unsaturated) heterocyclic ring having onenitrogen (wherein A is carbon, B is nitrogen, and C is CH—CH);

(3) a 6-membered aromatic (unsaturated) ring (wherein A is carbon, B isCH, and C is —CH—CH—);

or a pharmaceutically acceptable salt, or pro-drug form thereof.

Preferred compounds of this invention include:

(4-Thiophen-2-yl-phenyl)-(5H-10,11-dihydro-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone;

[4-(5-Bromo-thiophen-2-yl)-phenyl]-(5H-10,11-dihydro-pyrrolo[2,1c][1,4]benzodiazepin-10-yl)-methanone;

[2-Chloro-4-(5-chloro-thiophen-3-yl)-phenyl]-(5H-10,11-dihydro-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone;

(2-Chloro-4-thiophen-2-yl-phenyl)-(5H-10,11-dihydro-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone;

[2-Chloro-4-(5-chloro-thiophen-2-yl)-phenyl]-(5H-10,11-dihydro-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone;

[2-Chloro-4-(5-methyl-thiophen-2-yl)-phenyl]-(5H-10,11-dihydro-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone;

(2-Chloro-4-thiophen-3-yl-phenyl)-(5H-10,11-dihydro-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone;

[2-Chloro-4-(5-chloro-thiophen-3-yl)-phenyl]-(5H-10,11-dihydro-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone;

(2-Methyl-4-thiophen-2-yl-phenyl)-(5,11-dihydro-pyrido[2,3-b][1,5]benzodiazepin-10-yl)-methanone;and

[2-Chloro-4-(5-chloro-thiophen-2-yl)-phenyl]-(5,11-dihydro-pyrido[2,3-b][1,5]benzodiazepin-10-yl)-methanone.

(2-Chloro-4-thiophen-3-yl-phenyl)-(5,11-dihydro-pyrido[2,3-b][1,5]benzodiazepin-10-yl)-methanone.

It is understood by those in the art that some of the compounds of thisinvention depending on the definition of R¹, R², R³, and R⁴ may containone or more asymmetric centers and may thus give rise to optical isomersand diastereomers. The present invention includes such optical isomersand diastereomers; as well as the racemic and resolved, enantiomericallypure R and S stereoisomers and pharmaceutically acceptable saltsthereof, which possess the indicated activity. Optical isomers may beobtained in pure form by standard procedures known to those skilled inthe art. It is also understood that this invention encompasses allpossible regioisomers, and mixtures thereof which possess the indicatedactivity. Such regioisomers may be obtained in pure form by standardseparation procedures known to those skilled in the art.

Also according to the present invention there is provided a method oftreating or preventing disorders which are remedied or alleviated byvasopressin receptor agonist. Methods of this invention for inducingvasopressin agonism in a mammal include, but are not limited to, methodsof treating, alleviating or preventing diabetes insipidus, nocturnalenuresis, nocturia, urinary incontinence, bleeding and coagulationdisorders, and temporary delay of urination whenever desirable in humansor other mammals, which comprises administering to a human or othermammal an effective amount of a compound or a pharmaceutical compositionof the invention.

The present invention accordingly provides a pharmaceutical compositionwhich comprises a compound of this invention in combination orassociation with a pharmaceutically acceptable carrier. In particular,the present invention provides a pharmaceutical composition whichcomprises an effective amount of a compound of this invention and apharmaceutically acceptable carrier.

The compositions are preferably adapted for oral administration.However, they may be adapted for other modes of administration, forexample, parenteral administration for patients suffering fromcoagulation disorders.

In order to obtain consistency of administration, it is preferred that acomposition of the invention is in the form of a unit dose. Suitableunit dose forms include tablets, capsules and powders in sachets orvials. Such unit dose forms may contain from 0.1 to 1000 mg of acompound of the invention and preferably from 2 to 50 mg. Still furtherpreferred unit dosage forms contain 5 to 25 mg of a compound of thepresent invention. The compounds of the present invention can beadministered orally at a dose range of about 0.01 to 100 mg/kg orpreferably at a dose range of 0.1 to 10 mg/kg. Such compositions may beadministered from 1 to 6 times a day, more usually from 1 to 4 times aday. The compositions of the invention may be formulated withconventional excipients, such as a filler, a disintegrating agent, abinder, a lubricant, a flavoring agent and the like. They are formulatedin conventional manner, for example, in a manner similar to that usedfor known antihypertensive agents, diuretics and β-blocking agents.

Also according to the present invention there are provided processes forproducing the compounds of the present invention.

PROCESS OF THE INVENTION

The compounds of the present invention of general formula (I) mayconveniently be prepared according to the process shown in Scheme 1.

Thus, a tricyclic benzodiazepine of formula (1, wherein m, Y, A, B, C,R³and R⁴ are hereinbefore defined) is reacted with an appropriatelysubstituted acylating agent such as a haloaroyl halide, preferably abromoaroyl (iodoaroyl) chloride of formula (2, wherein J=COCl, X=Br orI, and R¹, R² are hereinbefore defined) in the presence of an inorganicbase such as potassium carbonate in a polar, aprotic solvent such asN,N-dimnethylformnamide; or an organic base in an aprotic solvent suchas dichioromethane or tetrahydrofuran at temperatures ranging from −40°C. to 50° C. to yield the intermediate acylated derivative (3).

Alternatively, the acylating species can be a mixed anhydride of thecorresponding carboxylic acid, such as that prepared treating said acidwith 2,4,6-trichlorobenzoyl chloride in an aprotic organic solvent suchas dichloromethane according to the procedure of Inanaga et al., BullChem. Soc. Jpn., 52, 1989 (1979). Treatment of said mixed anhydride ofgeneral formula (2) with the tricyclic benzodiazepine of formula (1) ina solvent such as dichloromethane and in the presence of an organic baseat temperatures ranging from 0° C. to the reflux temperatures of thesolvent, yields the intermediate acylated derivative (3) of Scheme (I)

The acylating intermediate of formula (2) is ultimately chosen on thebasis of its compatibility with the R¹, R², R³ and R⁴ groups, and itsreactivity with the tricyclic benzodiazepine of formula (1).

Reaction of a compound of formula (3, X=Br or I) with an appropriatelysubstituted thiophene boronic acid of formula (4, wherein R⁵ and R⁶ arehereinbefore defined, and W=B(OH)₂) in a mixture of solvents such astoluene-ethanol-water, and in the presence of a Pd(0) catalyst and abase such as sodium carbonate at temperatures ranging from ambient tothe reflux temperature of the solvent, yields the desired compounds offormula (I, wherein Y, m, A, B, C, R¹, R², R³, R⁴, R⁵ and R⁶ are asdefined above).

Alternatively, a compound of formula (3, X=I) is reacted with anappropriately substituted thiophene tri-alkyltin derivative of formula(4, wherein R⁵, and R⁶ are hereinbefore defined, and W=Sn(alkyl)₃) toafford the desired compound of formula (I, wherein Y, m, A, B, C, R¹,R², R³, R⁴, R⁵ and R⁶ are as defined above).

The preferred substituted 4-bromo(iodo) aroyl chlorides of formula (2)of Scheme I (X=Br or I, J=COCl) are either available commercially, orare known in the art, or can be readily prepared by procedures analogousto those in the literature for the known compounds.

The preferred substituted thiophene boronic acids of formula (4,W=B(OH)₂) are either available commercially, or or are known in the art,or can be readily prepared by procedures analogous to those in theliterature for the known compounds.

The preferred substituted thiophene tri-alkylstannanes of formula (4,W=Sn(alkyl)₃) of Scheme I are either available commercially, or can beconveniently prepared as shown in Scheme II. Thus, the correspondingbromo starting materials of formula (5, wherein W=Br, and R⁵ and R⁶ arehereinbefore defined) are first reacted with n-butyl lithium followed bytreatment of the intermediate lithiated species with a tri-alkyl(preferably tri-methyl or tri-n-butyl) tin chloride to give the desiredthiophene stannane intermediates (4, wherein W=Sn(alkyl)₃), and R⁵ andR⁶ are hereinbefore defined)

Alternatively, as shown in Scheme III, the bromo derivative (3) ofScheme I (wherein X=Br, and Y, m, A, B, C, R¹, R², R³ and R⁴ arehereinbefore defined) is reacted with an hexa-alkyl-di-tin in thepresence of a palladium (0) catalyst and lithium chloride to yield thetri-(alkyl)tin intermediate of formula (6). Further reaction of (6) withthe appropriately substituted thiophene halide of formula (5, whereinX=Br or I, and R⁵ and R⁶ are hereinbefore defined ) in the presence of apalladium(0) catalyst provides the desired compounds of formula (I) ofScheme I.

The desired compounds of formula (I) of Scheme I can be alternatively,prepared by the process shown in Scheme IV. Thus, an appropriatelysubstituted carboxylic acid derivative of formula (7, wherein P is acarboxylic acid protecting group, preferably P=alkyl or benzyl), isreacted with a thiophene tri-(alkyl)tin derivative (4) in the presenceof a palladium (0) catalyst to provide the intermediate ester (8).Subsequent unmasking of the carboxylic function followed by activationof the intermediate acid (9) using any of the procedures hereinbeforedescribed, and coupling of the intermediate (10) to the tricyclicbenzodiazepine of formula (1) provides the desired compounds (I) whereinY, m, A, B, C, R¹, R², R³, R⁴, R⁵ and R⁶ are as defined above.

Alternatively, the desired intermediates of formula (8) of Scheme IV canbe prepared from (7) and the thiophene boronic acid derivative offormula (4, wherein W=B(OH)₂) in a mixture of solvents such astoluene-ethanol-water, in the presence of a palladium(0) catalyst and abase such as sodium carbonate at temperatures ranging from ambient tothe reflux temperature of the solvent, as shown in Scheme V.

The compounds of formula (I) that are 5-substituted 2-alkylthiophenescan be conveniently prepared as shown in Scheme VI, by reaction of theintermediate bromide of formula (3) with a 2-alkyl thiophene of formula11 in the presence of a palladium(0) catalyst, potassium acetate and asolvent such as N,N-dimethyacetamide in a Carius tube at temperatures upto 150° C., essentially according to the procedure of Ohta et al.,Heterocycles, 31, 1951 (1990).

The subject compounds of the present invention were tested forbiological activity according to the following procedures.

Vasopressin V₂ Agonist Effects of Test Compounds in Normal ConsciousWater-Loaded Rats

Male or female normotensive Sprague-Dawley rats (Charles RiverLaboratories, Inc., Kingston, N.Y.) of 350-500 g body weight weresupplied with standard rodent diet (Purina Rodent Lab. Chow 5001) andwater ad libitum. On the day of test, rats were placed individually intometabolic cages equipped with devices to separate the feces from theurine and containers for collection of urine. A test compound or areference agent was given at an oral dose of 10 mg/Kg in a volume of 10mL/Kg. The vehicle used was 20% dimethylsulfoxide (DMSO) in 2.5%preboiled corn starch. Thirty minutes after dosing the test compound,rats were gavaged with water at 30 mL/Kg into the stomach using afeeding needle. During the test, rats were not provided with water orfood. Urine was collected for four hours after dosing of the testcompound. At the end of four hours, urine volume was measured. Urinaryosmolality was determined using a Fiske One-Ten Osmometer (FiskeAssociates, Norwood, Mass., 02062) or an Advanced CRYOMATIC Osmometer,Model 3C2 (Advanced Instruments, Norwood, Mass.). Determinations of Na⁺,K⁺ and Cl⁻ ion were carried out using ion specific electrodes in aBeckman SYNCBRON EL-ISE Electrolyte System analyzer. The urinaryosmolality should increase proportionally. In the screening test, tworats were used for each compound. If the difference in the urine volumeof the two rats was greater than 50%, a third rat was used.

The results of this study are shown in Table 1.

TABLE 1 Urine Volume Changes in Urinary Example (% decrease)^(a)Osmolality^(b) Rat Type^(c) 1 68 224 CD 2 74 819 CD 3 71 190 CD 4 74 365CD 5 63 180 CD 6 75 286 CD 7 75 282 CD 8 44 143 CD ^(a)Percent decreasein urine volume vs control at a dose of 10 mg/Kg ^(b)Percent changes inosmolality vs control at a dose of 10 mg/Kg ^(c)Rat model used:Sprague-Dawley (CD)

The following examples are presented to illustrate rather than limit thescope of the invention.

EXAMPLE 1(2-Chloro-4-thiophen-2-yl-phenyl)-(5H-10,11-dihydro-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone

Step A.(4-Bromo-2-chloro-phenyl)-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone

N,N-dimethylformamide (1 drop) was added to a solution of4-bromo-2-chlorobenzoic acid (2.30 g) in anhydrous tetrahydrofuran (20mL). Oxalyl chloride (1.46 g) was added and the mixture was warmed toreflux. The resultant solution was cooled to ambient temperature beforebeeing evaporated to dryness to give crude 4-bromo-2-chlorobenzoylchloride as a gold, viscous liquid, which was used without furtherpurification.

To a mixture of 10,11-dihydro-5H-pyrrolo[2,1-c][1,4]benzodiazepine (1.44g) and triethylamine (0.95 g) in dichloromethane (40 mL) cooled in anice bath, was added dropwise a solution of the crude4-bromo-2-chlorobenzoyl chloride (2.42 g) in dichloromethane (20 mL).The cooling bath was removed and after stirring for 22 hours, thereaction mixture was sequentially washed with water, saturated aqueoussodium bicarbonate, 0.5 N hydrochloric acid and water. Thedichloromethane solution was dried over anhydrous sodium sulfate,filtered and then evaporated to dryness to yield an off-white foam.Purification by flash chromatography on silica gel Merck-60 eluting withhexane-ethyl acetate (2:1) resulted in a white solid (3.02 g), m.p.77-80° C.

MS (EI, m/z); 400 [M]⁺.

Step B.(2-Chloro-4-thiophen-2-yl-phenyl)-(5H-10,11-dihydro-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone

Thiophene-2-boronic acid (0.51 g, 4 mmol) was added to a mixture of(4-bromo-2-chloro-phenyl)-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanoneof Step A (1.61 g, 4 mmol) and sodium carbonate (1.02 g, 9.6 mmol) intoluene (36 mL), ethanol (10 mL) and water (20 mL). The resultantsolution was purged with nitrogen for 15 minutes, and then tetrakis(triphenylphosphine) palladium(0) catalyst (0.18 g, 0.16 mmol) wasadded. The reaction mixture was heated to reflux for 17 hours, cooled toambient temperature, stirred for an additional 26 hours and filteredthrough Celite, which was then rinsed with ethyl acetate. The combinedfiltrate was diluted to 140 mL with water/ethyl acetate (1:1). The ethylacetate extracts were dried over anhydrous magnesium sulfate, filteredand evaporated to dryness. The residue (brown foam) was flashchromatographed on silica gel Merck-60 (eluant: hexane-ethyl acetate4:1) to yield an off-white foam, which was redissolved indichloromethane. Addition of hexane provided the title compound as awhite powder, m.p. 129.5-132° C.

¹H NMR (DMSO-d₆, 400 MHz): δ 5.28 (br m, 4H), 5.90 (t, 1H), 5.99 (s,1H), 6.82 (s, 1H), 7.10 (m, 4H), 7.40 (m, 3H), 7.60 (d, 2H), 7.64 (s,1H); MS (EI, m/z): 404 [M]⁺; Anal. Calcd. for C₂₃H₁₇ClN₂OS: C 68.22, H4.23, N 6.92. Found: C 68.30, H 4.26, N6.74;

EXAMPLE 2[2-Chloro-4-(5-chloro-thiphen-2-yl)-phenyl]-(5H-10,11-dihydropyrrolo[2,1-c][1.4]benzodiazepin-10-yl)-methanone

5-Chloro-thiophene-2-boronic acid (0.65 g, 4 mmol) was added to amixture of (4-bromo-2-chloro-phenyl)-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone of Example 1, StepA (1.61 g, 4 mmol) and sodium carbonate (1.02 g, 0.6 mmol) in toluene(36 mL), ethanol (10 mL) and water (20 mL). The resulting solution waspurged with nitrogen for 10 minutes, and then tetrakis(triphenylphosphine) palladium(0) catalyst (0.18 g, 0.16 mmol) wasadded. The solution was heated at reflux for 41 hours, cooled to ambienttemperature and filtered through Celite, which was rinsed with ethylacetate. The combined filtrate was diluted to 140 mL with water/ethylacetate (1:1), and the combined organic extracts were dried overanhydrous magnesium sulfate, filtered and evaporated to dryness. Theresidue (brown foam) was flash chromatographed on silica gel Merck-60(eluant: hexane-ethyl acetate 4:1) to yield a white foam which wasredissolved in dichloromethane. Addition of hexane provided the titlecompound as an off-white solid, m.p. 119.5-122° C.

¹H NMR (DMSO-d₆, 400 MHz): δ 5.28 (br m, 4H), 5.90 (t, 1H), 5.99 (s,1H), 6.81 (s, 1H), 7.10 (m, 4H), 7.40 (d, 3H), 7.50 (d, 1H), 7.64 (s,1H); MS (+FAB, m/z): 461 [M+Na]⁺, 439 [M+H]⁺; Anal. Calcd. forC₂₃H₁₆Cl₂N₂OS: C 62.88, H 3.67, N 6.38. Found: C 62.52, H 3.69, N 6.27.

EXAMPLE 3[2-Chloro-4-(5-methyl-thiophen-2-yl)-phenyl]-(5H-10,11-dihydro-pyrrolo[2,1-c][1.41]benzodiazepin-10-yl)-methanone

The title compound was prepared essentially according to the conditionsset forth by Ohta et al., Heterocycles, 31, 1951 (1990). Potassiumacetate (0.44 g, 4.5 mmol) was added to a solution of(4-bromo-2-chloro-phenyl)-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanoneof Example 1, Step A (1.2 g, 3 mmol) and 2-methylthiophene (1.5 mL,15.49 mmol) in N,N-dimethyl acetamide (7.5 mL), in a 15 mL Carius tube.The resultant solution was purged with nitrogen for 15 minutes, thentetrakis (triphenylphosphine) palladium(0) catalyst (0.17 g, 0.15 mmol)was added. The sealed tube was heated in an oil bath at 150° C. for 16.5hours. The solvent was removed in vacuo, and the residue was trituratedwith water (10 mL) and extracted with dichloromethane. The organicextracts were dried over anhydrous sodium sulfate, filtered andevaporated to yield a dark brown oil. Flash chromatography of theresidue on silica gel Merck-60 (eluant: hexane-ethyl acetate 2:1)resulted in an off-white foam which was redissolved in dichloromethane.Addition of hexane provided the title compound as an off-white powder,m.p. 154-155.5° C.

¹H NMR (DMSO-d₆, 400 MHz): δ 2.43 (s, 3H) 5.27 (br m, 4H), 5.90 (t, 1H),5.98 (s, 1H), 6.81 (m, 2H), 7.04 (m, 3H), 7.32 (s, 2H), 7.39 (d, 2H),7.55 (s, 1H); MS (EI, m/z): 418 [M]⁺; Anal. Calcd. for C₂₄H₁₉ClN₂OS: C68.81, H 4.57, N 6.69. Found: C 68.77, H 4.69, N 6.61.

EXAMPLE 4(2-Chloro-4-thiophen-3-yl-phenyl)-(5H-10,11-dihydro-pyrrolo[2,1-c][4,1]benzodiazepin-10-yl)-methanone

Thiophene-3-boronic acid (0.51 g, 4 mmol) was added to a mixture of(4-bromo-2-chloro-phenyl)-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl-methanoneof Example 1, Step A (1.61 g, 4 mmol) and sodium carbonate (1.02 g, 9.6mmol) in toluene (36 mL), ethanol (10 mL) and water (20 mL). Theresultant solution was purged with nitrogen for 10 minutes, thentetrakis (triphenylphosphine) palladium(0) catalyst (0.18 g, 0.16 mmol)was added. The reaction mixture was heated to reflux for 64 hours,cooled to ambient temperature, filtered through Celite, which was thenrinsed with ethyl acetate. The combined filtrate was diluted to 140 mLwith water-ethyl acetate (1:1), and the aqueous layer extracted withethyl acetate. The combined extracts were dried over anhydrous magnesiumsulfate, filtered and evaporated to dryness to yield a brown foam. Flashchromatography of the residue on silica gel Merck-60 (eluant:hexane-ethyl acetate 4:1) provided the title compound as a white solid,m.p.dec. 101° C.

¹H NMR (DMSO-d₆, 400 MHz): δ 5.28 (br m, 4H), 5.90 (t, 1H), 5.99 (s,1H), 6.81 (s, 1H), 7.03 (m, 3H), 7.37 (m, 2H), 7.55 (m, 3H), 7.74 (s,1H), 7.99 (s, 1H); MS (EI, m/z): 404 [M]⁺; Anal. Calcd. forC₂₃H₁₇ClN₂OS: C 68.23, H 4.23, N 6.92. Found: C 67.98, H 4.49, N 6.88

EXAMPLE 5[2-Chloro-4-(5-chloro-thiophen-3-yl-phenyl)-(5H-10,11-dihydro-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone

5-Bromo-2-chloro-thiophene (1.16 g, 5.79 mmol) was added to a mixture of3-chloro-4-carboxyboronic acid (1.09 g, 5.5 mmol), triethylamine (4 mL)and tetrakis (triphenylphosphine) palladium(0) catalyst (0.2 g, 0.17mmol) in pre-purged N,N-dimethylformamide (1 mL). The reaction mixturewas heated to reflux for 24 hours, the solvent was concentrated in vacuoand the residue was taken into water. The aqueous solution was washedwith ether and added to ice/concentrated hydrochloric acid (10 mL). Thewhite precipitate was extracted into dichloromethane and the solutionwashed with brine. The combined extracts were dried over anhydrousmagnesium sulfate, filtered, and evaporated to dryness to yield theexpected acid (0.65 g).

The freshly prepared acid (0.65 g, 2.38 mmol) was dissolved in anhydroustetrahydrofuran (20 mL). Oxalyl chloride (0.22 mL, 2.5 mmol) andN,N-dimethylformamide (1 drop) were added and the mixture was warmed to35° C. for 10 minutes. The resultant solution was cooled to ambienttemperature before being evaporated to dryness to give the crude acidchloride, which was used without further purification.

To a mixture of 10,11-dihydro-5H-pyrrolo[2,1-c][1,4]-benzodiazepine(0.417 g, 2.3 mmol) and diisopropylethylamine (0.5 mL, 2.8 mmol) indichloromethane (10 mL) was added a solution of the crude acid chloridein dichloromethane (2 mL). The reaction was stirred at room temperatureovernight. The organic solution was washed with 1N hydrochloric acid andbrine, dried over anhydrous sodium sulfate, filtered and evaporated todryness. Purification by flash chromatography on silica gel Merck-60(eluant: hexane-ethyl acetate 2:1) provided the title compound (0.1 g)as a white solid, m.p. 97-99° C.

¹H NMR (DMSO-d₆, 400 MHz): δ 5.00-5.35 (m, 4H), 5.91 (s, 1H), 5.99 (s,1H), 6.81 (s, 1H), 7.1 (m, 3H), 7.21-8.01 (m, 6H); MS (EI, m/z): 438,440, 442 [M]⁺;

EXAMPLE 6(2-Methyl-4-thiophen-2-yl-phenyl)-(5,11-dihydro-pyrido[2,3-b][1,5]benzodiazepin-10-yl)-methanone

Step A.(4-Bromo-2-methyl-phenyl)-(5,11-dihydro-pyrido[2,3-b][1,5]benzodiazepin-6-yl)-methanone

A suspension of 4-bromo-2-methylbenzoic acid (4.9 g, 22.8 mmol) indichloromethane containing a few drops of N,N-dimethylformamide wastreated dropwise under nitrogen with oxalyl chloride (2.4 mL, 27.5 mol).After gas evolution subsided, the reaction mixture was refluxed for anadditional 15 minutes and then evaporated to dryness in vacuo to providethe crude 4-bromo-2-methylbenzoyl chloride.

To a solution of 6,11-dihydro-5H-pyrido[2,3-b][1,5]benzodiazepine (3 g,15.2 mmol) in N,N-dimethylformamide under nitrogen was added solidpotassium carbonate (6.3 g, 45.6 mmol). A solution of the crude4-bromo-2-methylbenzoyl chloride (22.8 mmol) in N,N-dimethylformamnidewas added dropwise, and the mixture stirred at room temperature for 15minutes. Excess potassium carbonate was filtered off, the filtrate waswashed with water and the aqueous layer was extracted with chloroform.The extracts were dried over anhydrous sodium sulfate, evaporated todryness and the residue dissolved in dichloromethane and absorbed onto asilica Merck-60 flash column. Elution with 20% ethyl acetate in hexaneprovided the title compound (3.2 g) as a foam which crystallized uponsonication from ethanol/hexane.

¹H NMR (DMSO-d₆, 400 MHz): δ 2.04 (s, 3H), 4.10 and 5.46 (dd, 2H), 6.54(m, 1H), 6.68 (m, 1H), 6.78 (m, 1H), 6.90 (m, 1H), 7.00 (m, 1H),7.18-7.29 (m, 1H), 8.10 (m, 1H), 9.55 (s, 1H); MS (EI, m/z): 393/395[M]⁺; Anal. Calcd. for C₂₀H₁₆BrN₃O+0.05 C₂H₆O: C 60.89, H 4.13, N 10.61.Found: C 60.49, H 4.07, N 10.44.

Step B.(2-Methyl-4-thiophen-2-yl-phenyl)-(5,11-dihydro-pyrido[2,3-b][1,5]benzodiazepin-10-yl)-methanone

To a solution of(4-bromo-2-methyl-phenyl)-(5,11-dihydro-pyrido[2,3-b][1,5]benzodiazepin-6-yl)-methanone(0.5 g, 1.27 mmol) of Step A, thiophene-2-boronic acid (0.167 g, 1.30mmol) and sodium carbonate (0.595 g, 5.6 mmol) in toluene (20 mL),ethanol (10 mL) and water (10 mL) under nitrogen was added tetrakis(triphenylphosphine) palladium(0) catalyst (0.066 g, 0.057 mmol). Themixture was heated at reflux for 19 hours. Additional boronic acid(0.170 g, 1.30 mmol) and catalyst (0.050 g, 0.043 mmol) were added andreflux resumed for 3 hours. After stirring overnight at roomtemperature, the mixture was filtered through Celite and the cake washedwith ethyl acetate. The combined organic layers were washed with water,dried over anhydrous sodium sulfate, and evaporated to dryness. Theresidue was dissolved in dichloromethane, absorbed onto a silicaMerck-60 flash column and eluted with an ethyl acetate- hexane gradient(from 30 to 50%) to provide the title compound (0.29 g) as a foam, whichcrystallized as a white solid by trituration with diethyl ether/ hexane,m.p. 118-120° C.

¹H NMR (DMSO-d₆, 400 MHz): δ 2.11 (s, 3H), 4.10 and 5.49 (dd, 2H), 6.52(m, 1H), 6.70 (m, 1H), 6.90-7.00 (m, 2H), 7.08 (m, 1H), 7.21-7.30 (m,2H), 7.35 (m, 1H), 7.45 (m, 1H), 7.51 (m, 1H), 7.59 (m, 1H), 8.11 (m,1H), 9.56 (s, 1H); MS (EI, m/z): 397 [M]⁺; Anal. Calcd. for C₂₄H₁₉N₃OS:C 72.52, H 4.82, N 10.57. Found: C 72.79, H 5.18, N 10.52.

EXAMPLE 7[2-Chloro-4-(5-chloro-thiphen-2-yl-phenyl]-(5,11-dihydro-pyrido[2,3-b][1,5]benzodiazepin-10-yl)-methanone

Step A.(4-Bromo-2-chloro-phenyl)-(5,11-dihydro-pyrido[2,3-b][1,5]benzodiazepin-10-yl)-methanone

A suspension of 4-bromo-2-chlorobenzoic acid (5.4 g, 22.9 mmol) indichloromethane (40 mL) containing a few drops of N,N-dimethylformamidewas treated dropwise under nitrogen with oxalyl chloride (2.4 mL, 27.5mmol). After the gas evolution subsided, the reaction mixture wasrefluxed for an additional 15 minutes and evaporated to dryness in vacuoto provide the crude acid chloride.

To a solution of 6,11-dihydro-5H-pyrido[2,3-b][1,5]benzodiazepine (3 g,15.2 mmol) in N,N-dimethylformamide under nitrogen were added solidpotassium carbonate (6.3 g, 45.6 mmol). The mixture was treated dropwisewith a solution of the crude 4-bromo-2-chlorobenzoyl chloride (22.9mmol) in N,N-dimethylformamide. After stirring at room temperature for15 minutes, water was added under stirring. The resulting solid wascollected, dissolved in chloroform, and the solution washed with 1N NaOHand brine. The organic layer was dried over anhydrous sodium sulfate andevaporated to dryness. The residue (purple foam) was dissolved indichloromethane and absorbed onto a silica Merck-60 flash column.Elution with 20% ethyl acetate in hexane provided 3.4 g of the titlecompound as a foam which crystallized as a white solid by triturationfrom ethanol-ether, m.p. 165-168° C.

¹H NMR (DMSO-d6, 400 MHz): δ 4.13 and 5.42 (dd, 2H), 6.54 (m, IH),6.73-6.79 (m, 2H), 7.01 (m, 1H), 7.22-7.34 (m, 2H), 7.45 (m, 1H),7.48-7.62 (m, 2H), 8.10 (m, 1H), 9.55 (s, 1H); MS (EI, m/z): 413/415/417[M]⁺; Anal. Calcd. for C₁₉H₁₃BrClN₃O: C 55.03, H 3.16, N 10.13. Found: C54.81, H 3.15, N 9.86.

Step B. [2-Chloro-4-(5-chloro-thiphen-2-yl)-phenyl]-(5,11-dihydro-pyrido[2,3-b][1,5]benzodiazepin-10-yl)-methanone 0.28 solvate with hexane

To a solution of the(4-bromo-2-chloro-phenyl)-(5,11-dihydro-pyrido[2,3-b][1,5]benzodiazepin-10-yl)-methanoneof Step A (0.5 g, 1.2 mmol), 5-chlorothiophene-2-boronic acid (0.21 g,1.29 mmol) and sodium carbonate (0.57 g, 5.38 mmol) in toluene (20 mL),ethanol (10 mL) and water (10 mL) under nitrogen was added tetrakis(triphenylphosphine) palladium(0) catalyst (0.06 g, 0.052 mmol). Themixture was heated at reflux for 18 hours. Additional boronic acid (0.2g, 1.29 mmol) and catalyst (0.065 g, 0.056 mmol) were added. After 5hours the mixture was cooled and filtered through Celite which was thenrinsed with ethyl acetate. The organic layer was washed with water,dried over anhydous sodium sulfate, and evaporated to dryness. Theresidue was dissolved in dichloromethane and absorbed onto a column offlash silica gel Merck-60. Elution with 25% ethyl acetate in hexaneprovided the title compound (0.25 g) as a foam which crystallized as apale yellow solid by trituration from ethanol-hexane, m.p. 131-134° C.

¹H NMR (DMSO-d₆, 400 MHz): δ 4.14 and 5.44 (dd, 2H), 6.52 (m, 1H),6.72-6.79 (m, 2H), 7.00 (m, 1H), 7.15 (m, 1H), 7.22-7.26 (m, 2H),7.41-7.48 (m, 2H), 7.53-7.63 (m, 2H), 8.11 (m, 1H), 9.56 (s, 1H); MS(EI, m/z): 451/453/455 [M]⁺; Anal. Calcd. for C₂₃H₁₅Cl₂N₃OS+0.28 C₆H₁₄:C 62.21, H 4.00, N 8.82. Found: C 62.07, H 3.98, N 8.96.

EXAMPLE 8(2-Chloro-4-thiophen-3-yl-phenyl)-(5,11-dihydro-pyrido[2,3-b][1,5]benzodiazepin-10-yl)-methanone

To a solution of the(4-bromo-2-chloro-phenyl)-(5,11-dihydro-pyrido[2,3-b][1,5]benzodiazepin-10-yl)-methanoneof Example 7, Step A (0.5 g, 1.23 mmol), thiophene-3-boronic acid (0.158g, 1.23 mmol) and sodium carbonate (0.568 g, 5.36 mmol) in toluene (20mL), ethanol (10 mL) and water (10 mL) under nitrogen was added thetetrakis (triphenylphosphine) palladium(0) catalyst (0.06 g, 0.052mmol). The reaction was heated at reflux for 20 hours, cooled, andfiltered through Celite, which was then rinsed with ethyl acetate. Theorganic layer was washed with water, dried over sodium sulfate andevaporated to dryness. The residue (blue-green oil) was dissolved indichloromethane and absorbed onto a silica Merck-60 flash column.Elution with 25% ethyl acetate in hexane yielded a foam (0.24 g) whichwas triturated with diethyl ether-hexane to provide the title compoundas a white solid, m.p. 125-130° C.

¹H NMR (DMSO-d₆, 400 MHz): 4.14 and 5.45 (dd, 2H), 6.51 (m, 1H),6.76-6.80 (m, 2H), 6.99 (m, 1H), 7.22-7.26 (m, 2H), 7.54-7.71 (m, 5H),7.98 (m, 1H), 8.11 (m, 1H), 9.56 (s, 1H); MS (EI, m/z): 417/419 [M]⁺;Anal. Calcd. for C₂₃H₁₆ClN₃OS: C 66.10, H 3.86, N 10.05. Found: C 66.38,H 4.11, N9.85.

What is claimed:
 1. A compound of the formula:

wherein: R¹, R², R⁵ and R⁶ are independently, selected from hydrogen,lower alkyl (C₁-C₆), lower alkoxy (C₁-C₆), halogen, and CF₃; R₃ and R₄are independently, selected from the group comprising hydrogen, loweralkyl (C₁-C₆), halogen, amino, (C₁-C₆) lower alkoxy, or (C₁-C6) loweralkylamino; or a pharmaceutically acceptable salt thereof.
 2. A compoundaccording to claim 1 which is(2-methyl-4-thiophen-2-yl-phenyl)-(5,11-dihydro-pyrido[2,3-b][1,5]benzodiazepin-10-yl)-methanone,or a pharmaceutically acceptable salt thereof.
 3. A compound accordingto claim 1 which is[2-chloro-4-(5-chloro-thiophen-2-yl)-phenyl]-(5,11-dihydro-pyrido[2,3-b][1,5]benzodiazepin-10-yl)-methanone,or a pharmaceutically acceptable salt thereof.
 4. A compound accordingto claim 1 which is(2-chloro-4-thiophen-3-yl-phenyl)-(5,11-dihydro-pyrido[2,3-b][1,5]benzodiazepin-10-yl)-methanone,or a pharmaceutically acceptable salt thereof.
 5. A pharmaceuticalcomposition comprising a pharmaceutically effective amount of a compoundof claim 1 and a pharmaceutically acceptable carrier.
 6. A method oftreating in a mammal disorders which are remedied or alleviated byvasopressin agonist activity selected from the group of diabetesinsipidus, nocturnal enuresis, nocturia, urinary incontinence, orbleeding or coagulation disorders, the method comprising administeringto a mammal in need thereof a pharmaceutically effective amount of acompound of claim 1, or a pharmaceutically acceptable salt form thereof.7. A method for inducing temporary delay of urination in a mammal, themethod comprising administering to a mammal need thereof apharmaceutically effective amount of a compound of claim 1, or apharmaceutically acceptable salt thereof.